ANTIGEN PRESENTATION AND PROCESSING; MECHANISMS OF LYMPHOCYTE ACTIVATION

Experiments have made it clear that for T cells to respond to antigen, they had to encounter not only the antigen itself, but the antigen along with particular MHC proteins T cells respond to antigen only if the antigen is presented to them by different kinds of cells-called antigen-presenting cells. Professional antigen presenting cells (APC):
Monocytes/macrophages Dendritic cells B cells

B cells and T cells recognize different substances as antigens and in a different form. The B cell uses cell surface-bound immunoglobulin as a receptor and the specificity of that receptor is the same as the immunoglobulin that it is able to secrete after activation.

B cells recognize the following antigens in soluble form Proteins (both conformational determinants and determinants exposed by denaturation or proteolysis)
Nucleic acids Polysaccharides Some lipids Small chemicals (haptens)

In contrast, the majority of antigens for T cells are proteins, and these must be fragmented and recognized in association with MHC products expressed on the surface of nucleated cells, not in a soluble form. T cells are grouped functionally according to the class of MHC molecules that associate with the peptide fragments of the protein:
Helper T cells (CD4) recognize only those peptides associated with class II MHC molecules. Cytotoxic T cells (CD8)recognize only those peptides associated with class I MHC molecules.

Histocompatibility (transplantation) antigens Antigens on tissues and cells that determine their rejection when grafted between two genetically different individuals Major histocompatibility (MHC) antigens Histocompatibility antigens that cause a very strong immune response and are most important in rejection MHC complex Group of genes on a single chromosome encoding the MHC antigens (chromosome 6 in man)

HLA (human leukocyte antigens):

MHC antigens of man (first detected on leukocytes)

H-2 antigens:
MHC antigens of mouse

AG processing is a generation of peptides from intact proteins AG presentation is a display of the AG peptides in complex with MHC molecules on the surface of the cell

In humans, the MHC gene products are termed human leukocyte antigens (HLA), and their corresponding genes are found on the short arm of chromosome 6. The HLA are glycoproteins present on cell surfaces that enable T cells to recognize and bind antigenic peptides. They function in immune recognition.

The MHC is a collection of highly polymorphic genes encoding the proteins that regulate immune responses.
These genes include, the class I (express in all nucleated cells) and class II (express only in APC) cell surface proteins and the-class III genes that encode some soluble complement molecules and cytokines.

MHC products play an important role in antigen recognition by T cells.

The class I and class II antigens are expressed on cells and tissues. Whereas as class III antigens are represented on proteins in serum and other body fluids (e.g.C4, C2, factor B, TNF). Antigens of class III gene products have no role in graft rejection.

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In vertebrates it has a major influence on graft survival. Individuals identical for this region can exchange grafts more successfully than MHC non-identical combinations. There also exist many "minor" histocompatibility loci which also influence graft survival but which are individually weak.

The MHC complex contains a number of genes that control several antigens, most of which influence allograft rejection. MHC class I and II loci have many more than 100 different alleles in mice and in humans. These genes, and therefore, these proteins, vary much more from individual to individual than any other known protein

All class I molecules share common structures. These include: 1. An a heavy chain
a. It has three extracellular domains, two of which form the peptide-binding groove. b. The carboxy terminus lies within the cytoplasm.

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2. A single transmembrane domain 3. A light chain, called 2-microglobin

a. It is not encoded by a gene in the MHC region. b. It functions to transport MHC class I molecules to the cell surface

The class I gene complex contains three major loci, B, C and A and other undefined minor loci. Each major locus codes for a polypeptide; the alpha-chain that contains antigenic determinants, is polymorphic (has many alleles). It associates with beta-2 microglobulin (beta-chain), encoded by a gene outside the MHC complex, and expressed on the cell surface.

An chain with three extracellular domains Single transmembrane domain Separately encoded chain ( 2 microglobulin)

HLA class II proteins are expressed on a more restricted set of cells, including antigen presenting cells (dendritic cells, Langerhans cells, activated macrophages, B cells, activated T cells, and activated endothelial cells). These proteins bind exogenous peptide epitopes that have been endocytosed and processed and are necessary for antigen recognition by CD4+ helper T lymphocytes.

Class II genes code for cell-surface glycoproteins with two polypeptide components called and . In humans, the class II genes include HLA-DR, HLA-DQ, and HLA-DP. As with the class I system, the class II genes are highly polymorphic; there are approximately 500 total class II gene products. They are also co dominantly expressed.

The three loci, DP, DQ and DR; each codes for one alpha- and one beta-chain polypeptide which associate together to form the class II antigens. Like the class I antigens, the class II antigens are also polymorphic. The DR locus may contain more than one, possibly four, functional beta-chain genes.

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Internalized and digested exogenous antigens the resultant peptide fragments complex with class II molecules to be presented On the cell surface. Both and chains with two extracellular domains. Two transmembrane domains recognized by CD4+ helper T cells.

Antigen processing and presentation are processes that occur within a cell that result in fragmentation (proteolysis) of proteins, association of the fragments with MHC molecules, and expression of the peptide-MHC molecules at the cell surface where they can be recognized by the T cell receptor on a T cell. However, the path leading to the association of protein fragments with MHC molecules differs for class I and class II MHC.

The 2 classes of MHC molecule are specialized to present different sources of antigen. MHC class I molecules present endogenously synthesized antigens, e.g. viral proteins. MHC class II molecules present exogenously derived proteins, e.g. bacterial products or viral capsid proteins.

Proteins are fragmented in the cytosol by proteosomes (a complex of proteins having proteolytic activity) or by other proteases. The fragments are then transported across the membrane of the endoplasmic reticulum by transporter proteins. (The transporter proteins and some components of the proteosome have their genes in the MHC complex).

Specifically transported across the ER membrane by a heterodimeric transporter made up of the TAP1 and TAP2 molecule

Synthesis and assembly of class I heavy chain and beta2 microglobulin occurs in the endoplasmic reticulum. Within the endoplasmic reticulum, the MHC class I heavy chain, beta2microglobulin and peptide form a stable complex that is transported to the cell surface.

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Cytotoxic CD8+ve T lymphocytes recognize the peptides that are associated with the class I molecules. Thus the ability of a CD8+ cytotoxic T cell to recognize antigen depends on association of the antigen with a class I protein. This is termed MHC restriction. A cytotoxic T cell that kills a virus-infected cell will not kill a cell infected with the same virus if the cell does not also express the appropriate class I protein.

Only a limited group of cells express class II MHC, which includes the antigen presenting cells (APC). The expression of class II MHC molecules is either constitutive or inducible, especially by interferon-gamma in the case of macrophages.

Proteins from outside these antigen-presenting cells are nonspecifically internalized by endocytosis. In the endocytic vacuoles in these cells, enzymes degrade (process) the proteins into small(about 10-20amino acid) peptides.

The alpha and beta chains of MHC class II, along with an invariant chain, are synthesized, assembled in the endoplasmic reticulum, and transported through the Golgi and trans-Golgi apparatus to reach the endosome, where the invariant chain is digested, and the peptide fragments from the exogenous protein are able to associate with the class II MHC molecules, which are finally transported to the cell surface.

The invariant chain blocks access to the peptide-binding groove of the MHC class II molecule as it is transported through the cell. Once the vesicle containing the MHC class II molecule fuses with the endocytic vacuole containing the digested, internalized peptides, the invariant chain is released and degraded, leaving the MHC class II molecule accessible to the peptides in the vacuole. The MHC class II-peptide complex is then transported to the cell surface, where it can be recognized by helper T cells, which have antigen receptors that recognize only peptides bound to MHC class II molecules. This is termed MHC restriction.